Bioorthogonally Cross-Linked Hyaluronan-Laminin Hydrogels for 3D Neuronal Cell Culture and Biofabrication.
Michael JuryIsabelle MatthiesenFatemeh Rasti BoroojeniSaskia L LudwigLivia CivitelliThomas E WinklerRobert SelegårdAnna HerlandDaniel AiliPublished in: Advanced healthcare materials (2022)
Laminins (LNs) are key components in the extracellular matrix of neuronal tissues in the developing brain and neural stem cell niches. LN-presenting hydrogels can provide a biologically relevant matrix for the 3D culture of neurons toward development of advanced tissue models and cell-based therapies for the treatment of neurological disorders. Biologically derived hydrogels are rich in fragmented LN and are poorly defined concerning composition, which hampers clinical translation. Engineered hydrogels require elaborate and often cytotoxic chemistries for cross-linking and LN conjugation and provide limited possibilities to tailor the properties of the materials. Here a modular hydrogel system for neural 3D cell cultures, based on hyaluronan and poly(ethylene glycol), that is cross-linked and functionalized with human recombinant LN-521 using bioorthogonal copper-free click chemistry, is shown. Encapsulated human neuroblastoma cells demonstrate high viability and grow into spheroids. Long-term neuroepithelial stem cells (lt-NES) cultured in the hydrogels can undergo spontaneous differentiation to neural fate and demonstrate significantly higher viability than cells cultured without LN. The hydrogels further support the structural integrity of 3D bioprinted structures and maintain high viability of bioprinted and syringe extruded lt-NES, which can facilitate biofabrication and development of cell-based therapies.
Keyphrases
- extracellular matrix
- stem cells
- drug delivery
- hyaluronic acid
- endothelial cells
- tissue engineering
- drug release
- cell therapy
- wound healing
- single cell
- induced apoptosis
- cell cycle arrest
- cerebral ischemia
- gene expression
- spinal cord
- endoplasmic reticulum stress
- white matter
- multiple sclerosis
- mesenchymal stem cells
- mass spectrometry
- bone marrow
- induced pluripotent stem cells
- case report
- cell death
- spinal cord injury
- molecularly imprinted
- solid phase extraction